基于UPLC-Q-Orbitrap MS技术的沙棘黄酮类成分分析

利用UPLC-Q-Orbitrap MS技术对沙棘(Hippophae rhamnoides L.)果实中黄酮类物质进行系统剖析,并对不同产地沙棘果实中黄酮类化合物进行比较分析,为中药沙棘的化学物质基础和资源利用研究奠定基础。根据保留时间比对,精确分子量匹配和MS/MS碎片信息分析,共鉴定出58个化合物,包括34个黄酮醇、4个黄酮、14个花色素、1个二氢黄烷酮、2个二氢黄烷醇、2个黄烷醇和1个二氢查尔酮,其中14个花色素类化合物为首次在沙棘果实中鉴定。最终筛选得到38种差异积累的黄酮类化合物,其中大部分在青海和西藏产地的沙棘果中呈显著积累。研究丰富了沙棘果中黄酮类化合物化学物质基础研究,揭示了不同产地沙棘果中黄酮类化合物的组成及差异。

Flavonoids and Amino Acids Analysis of the Fruits of Cornus mas L. Introduced in Uzbekistan

Cornelian cherry is used in the food and pharmaceutical industry as an ornament, in traditional medicine, and in the manufacture of household items. It is widely used in medicine for the prevention and treatment of many diseases. Therefore, it is important to research the chemical composition of these species. In the article, based on our research, the analysis of the quantitative calculation of flavonoids and amino acids of the fruits of the “Elegant” and “Svetlyachok” cherry varieties was carried out. It was found that the amount of amino acids in the fruits of Cornus mas L., introduced in Uzbekistan, is higher in the variety “Svetlyachok” 2.643235 mg/g. In “Elegant” variety it was 1.794235 mg/g. The amount of 4 different flavonoids in the fruit was also determined. It has been established that the Elegant variety has a high concentration of lutein and rutin, and the svetlyachok has a high concentration of apigenin and quercetin.

基于UPLC特征图谱和多成分定量评价不同采收时间枸橘药材质量

目的:基于不同采收时间枸橘药材UPLC特征图谱,结合黄酮类和香豆素类成分的含量测定,明确枸橘成分积累规律,为枸橘的采收和质量控制提供依据。方法:采用UPLC法构建30批枸橘特征图谱,采用相似度评价、聚类分析、主成分分析和正交偏最小二乘判别分析对数据进行处理。采用HPLC法测定其中6个成分的含量。结果:30批样品UPLC特征图谱共标定25个共有峰,相似度为0.913~0.993,指认了7个成分,其中6个成分芸香柚皮苷、柚皮苷、枸橘苷、欧前胡素、异欧前胡素、葡萄内酯的含量分别为0.62~2.40 mg/g、5.75~27.52 mg/g、7.00~53.86 mg/g、0.63~2.94 mg/g、0.11~0.45 mg/g、3.69~9.29 mg/g;聚类分析可将样品分为3类,正交偏最小二乘判别分析筛选出5个质量差异标志性成分。不同采收时间枸橘药材可以通过果皮颜色区分,枸橘苷、柚皮苷、欧前胡素等是导致枸橘药材质量产生差异的主要标志物。结论:果皮颜色可作为枸橘药材的采收依据;枸橘苷、柚皮苷和欧前胡素等是枸橘药材质量差异标志物。

Ecological Analysis of Leontice L. Species Distributed in the Navoi Region, Kyzyl-Kum Desert (Uzbekistan)

Leontice L. is a perennial herbaceous plant belonging to the Berberidaceae family. Our research was conducted in Navoi region. The article presents an ecological analysis of Leontice L. species in the Navoi region. The genus Leontice grows mainly in mountainous areas and some are distributed in arid climates. According to the information that 2 species have been identified in Navoi region, these species are plants with flavanoids healing properties. Leontice species have been used in traditional medicine since ancient times in ethnobotany. The presence of this species was discovered during the research and Leontice has identified numerous unique compounds, such as alkaloids, flavonoids, and phenolic acids, with potential medicinal properties such as anti-inflammatory, antitumor, and antiviral effects. According to ecological analysis, the Leontice genus thrives on stony, gravelly, and fine-grained mountain slopes and ridges, foothills, mountains, rocks, colorful rock outcrops, rocky slopes, and sandy, clay, and gravel deserts.

Assessment of Some Secondary Metabolites, Minerals and Alcohol Content of Noni Juice Obtained by Fermentation of Morinda citrifolia L. Fruit from Senegal

The fruit of Morinda citrifolia L., commonly known as noni, has an extensive history of use as a food and traditional medicine around the world. Adding value to Morinda citrifolia L. products, particularly the fruit, could be one way of building resilience in vulnerable farming households. The aim of this study was to determine the secondary metabolite and mineral composition of noni juice obtained by fermenting the fruit of Morinda citrifolia L. Fruits were collected in August 2022 from the local field in Thiès region, West of Senegal. Extraction yields were determined and the secondary metabolites were determined using conventional analytical methods. Calcium, magnesium, iron, sodium and potassium were determined by atomic absorption spectrophotometer coupled with a CCD detector. The results show that an average fruit mass (503.2 ± 110.96 g) consists of 171.44 ± 50.01 g pulp and 34.06 ± 10.35 g seeds. The traditional extraction yield of noni juice is 16.46% after three weeks of fermentation. The contents of total polyphenols, flavonoids and tannins obtained in noni are 608.97 ± 4.53 mg EAG/100mL, 7.78 ± 0.01 mg EQ/100mL and 0.191 ± 0.01 mg EC/100mL respectively. The ethanol content of noni varies from 3.57 to 5.23 mL/100mL during extraction. Noni has a high calcium content with a concentration of 383.79 ± 33.23 mg/L. This is followed by a good concentration of magnesium, potassium and sodium, at 278.47 ± 26.30, 187.43 ± 10.7 and 155.95 ± 28.66 mg/L respectively. Noni also has an iron content of 202.15 ± 0.05 mg/L.

Phytochemical Screening and Assays of Phenolic Compounds in Senna occidentalis L. Leaf and Seed Extracts

Phytochemical screening and assay of secondary metabolites, crude extracts with distilled water, aqueous methanol, aqueous acetone and aqueous ethanol of leaves and seeds of Senna occidentalis L. were studied in this work. The aim was to verify the distribution of secondary metabolites according to S. occidentalis organs. Four leaf samples from four different localities (South, East, West and Central Senegal) and a mixed sample of seeds were used. Functional molecules such as polyphenols, flavonoids and tannins were then assessed in the leaves and seeds using various standard methods. The results show that Senna occidentalis L. leaf and seed samples display an identical and homogeneous profile, regardless of locality. They contain secondary metabolites and the polyphenol content of extracts from southern, eastern, western and central leaves is: 0.620 - 0.539 - 0.811 - 0.573 g GAE/100 g DM;flavonoids: 0.064 - 0.074 - 0.130 - 0.101 g CE/100 g DM and tannins: 0.326 - 0.264 - 0.269 - 0.494 g TAE/100 g DM. The efficacy of S. occidentalis L. infusions in therapy is thus justified by the presence of these metabolites, whose biological properties are well known. It is then possible to explore isolation of active principles of Senna occidentalis L. leaves and even seeds for producing medicines.

Citrus aurantium L. extract alleviate depression by inhibiting gut microbiota-mediated inflammation in mice

Gut microbiota plays a crucial role in the pathophysiology of depression.This study aimed to explore the antidepressant effect of mature whole Citrus aurantium fruit extract(FEMC)in the chronic unpredictable mild stress(CUMS)model.The behavioral tests were applied to assess antidepressant effect and 16S rRNA sequencing was used to analyze the changes of gut microbiota.The results showed that the major components of FEMC were naringin and neohesperidin and significantly increased the sucrose preference index of the mice.FEMC also could reduce the feeding latency in an open field test and the rest time in a novelty suppressed feeding test.In addition,FEMC could increase CUMS-induced reduction in the levels of BDNF,PSD95,and SYN in the hippocampus.Moreover,FEMC intervention slightly decreased the ratio of Firmicutes to Bacteroidota.Meanwhile,FEMC reduced the abundance of the Prevotellaceae_Ga6A1_group,[Ruminococcus]_torques_group,which have been reported to be closely related to inflammation.Bioinformatics analysis revealed that mitogen-activated protein kinase(MAPK)signaling pathway and lipopolysaccharide biosynthesis were involved in the anti-inflammatory effect of FEMC in the CUMS animal model.Finally,the ELISA results showed that FEMC could significantly reduce the expression of pro-inflammatory cytokines IL-6 and TNF-αin the serum of depressive mice.Our results suggest FEMC can am eliorate depressive behavior by i nhibiting gut microbiota-mediated inflammation in mice.

Phenolic and flavonoid contents of mandarin (Citrus reticulata Blanco) fruit tissues and their antioxidant capacity as evaluated by DPPH and ABTS methods

The total phenolic and flavonoid contents in the fruit tissues （peels, pulp residues, seeds, and juices） of 19 citrus genotypes belonged to Citrus reticulata Blanco were evaluated and their antioxidant capacity was tested by 2,2-diphenyl-l-picrylhydra- zyl radicals （DPPH） method and 2,2＇-azino-bis（3-ethylbenzthiozoline-6）-sulphonic acid （ABTS） method. The total phenolic and flavonoid contents, and their antioxidant capacity varied in different citrus fruit tissues. Generally, the peel had both the highest average of total phenolics （27.18 mg gallic acid equivalent （GAE） g^-1 DW） and total flavonoids （38.97 mg rutin equivalent （RE） g^-1 DW）. The highest antioxidant capacity was also the average of DPPH value （21.92 mg vitamin C equiv- alent antioxidant capacity （VCEAC） g^-1 DW） and average of ABTS value （78.70 mg VCEAC g-1 DW） in peel. The correlation coefficient between the total phenolics and their antioxidant capacity of different citrus fruits tissues ranged from 0.079 to 0.792, and from -0.150 to 0.664 for the total flavonoids. The antioxidant capacity of fruit tissues were correlated with the total phenoilc content and flavonoid content except in case of the peel. In addition, the total phenolic content and antioxidant capacity varied in different citrus genotypes. Manju and Karamandarin were better genotypes with higher antioxidation and the phenolic content, however Shagan was the poorest genotype with lower antioxidation and the phenolic content.

An experimental study exploring the influencing factors for ultrasonic-assisted extraction of flavonoid compounds from leaves of Amorpha fruticosa L.

In this paper,the ultrasonic-assisted extraction process of flavonoid compounds from leaves of Amorpha fruticosa is optimized.In single factor experiments,solid/liquid ratios,ultrasonic power,ethanol concentrations and extraction cycles were experimental factors.Box–Behnken central composite design and RSM analyzed the effects of the four factors on the yield of total flavonoids.The optimal extraction parameters were solid/liquid ratio 1:50 g/mL,ultrasonic power 316 W,ethanol concentration 50%,4 extraction cycles.In the optimized condition,the estimated value of the regression model was 66.6456 mg/g while the measured value was 66.4329 mg/g.

A New Biflavonoid from Stellera chamaejasme L.

A new biflavonoid, stelleranol, was isolated from the roots of Stellera chamaejasme L.. Its structure was determined by the analysis of MS and NMR data, especially 2D NMR spectra.

Effects of Total Flavonoids ofHippophae RhamnoidesL.on Intracellular Free Calciumin Cultured Vascular Smooth Muscle Cells of Spontaneously Hypertensive Rats and Wistar-Kyoto Rats

To explore the effects of total flavonoids of Hippophae rhamnoides L. （TFH） quercetin （Que） and isorhamnetin （Isor） on the intracellular free calcium （[Ca^2＋]） in vascular smooth muscle cells （VSMC） of spontaneously hypertensive rats （SHR） and Wistar-Kyoto rats （WKY）. Metheds： Fluo 3-acetoxymethylester（Fluo-3/AM） was used to observe the effects of TFH （100mg/L） and its essential monomers, namely Que （10^-4mol/L） and Isor （10^-4mol/L） on changes of [Ca^2＋]1 in cultured SHR and WKY VSMC （abbr. to Ca-SHR ＆ Ca-WKY） following exposure to high K^＋, norepinephrine （NE） and angiotensin Ⅱ （AngⅡ）, and to compare with the effects of verapamil （Ver）. Results： （1） TFH, Que and Isor had inhibitory effects on resting Ca-SHR （P〈0.05）, but had no significant effects on Ca-WKY （P〉0.05）. （2） High K^＋ could increase Ca-SHR more significantly than Ca-WKY （P〈0.05）; TFH, Que and Isor could inhibit the elevation of [Ca^2＋]1 induced by high K^＋ -depolarization, with the effects similar to that of Ver, and the effect on Ca-SHR was more significant than that on Ca-WKY （P〈0.05）. （3） NE and Ang Ⅱ could increase Ca-SHR more significantly than Ca-WKY （P〈0.05）, TFH, Que and Isor had remarkably inhibitory effect on the elevation of Ca-SHR and Ca-WKY induced by NE or Ang Ⅱ. （4） In the absence of extracellular Ca^2＋ , TFH, Que and Isor also had certain inhibitory effect on Ca-SHR and Ca-WKY induced by NE, and the effect on the former was more significant than that on the latter（P〈0.05）. Ceaclusiea： TFH, Que and Isor might decrease the levels of [Ca^2＋], in VSMCs by blocking both voltage-dependent calcium channels （VDC） and receptoroperated calcium channels （ROC） in physiological or pathological state, which may be one of the important mechanisms of their hypotensive and protective effects on target organs in patients with hypertension.

Quantification and analysis of anthocyanin and flavonoids compositions,and antioxidant activities in onions with three different colors

Onion（Allium cepa L.） is a very important vegetable crop widely cultivated throughout the world. In this study, we measured total anthocyanins and flavonols of onions of three colors. The fresh weight（FW） of total anthocyanins were（29.99±1.19）,（9.64±1.30） and（0.75±0.40） mg 100 g^–1 fruit in red, yellow and white onions, respectively. Likewise, the FW of total flavonoids were（111.10±5.98）,（36.64±3.59）, and 0 mg 100 g^–1 in red, yellow and white, respectively. Four types of anthocyanins（delphinidin 3,5-diglycosides, cyanidin 3,5-diglycosides, cyanidin 3-glycosides and cyanidin 3-（6′′-malonyl）-glucopyranoside） and two kinds of flavonoids（quercetin and quercetin 3-glycosides） were identified in two varieties（red and yellow bulb） of onions by HPLC/DAD-ESI/MS. The total polyphenol contents were also measured by means of Folin-Ciocalteu method. Moreover, all the concentrations of anthocyanins, flavonoids and polyphenols showed significantly positive correlations with antioxidant activities measured by DPPH˙, ABTS˙＋ and FRAP assays. This study provided information on anthocyanin and flavonoids compositions that will be useful for onion breeding.

Study of total phenol,flavonoids contents and phytochemical screening of various leaves crude extracts of locally grown Thymus vulgaris

Objective:To prepare various crude extracts using different polarities of solvent and to quantitatively evaluate their total phenol,flavonoids contents and phytochemical screening of Thymus vulgaris collected from Al Jabal Al Akhdar,Nizwa.Sultanate of Oman.Methods:The leave sample was extracted with methanol and evaporated.Then it was defatted with water and extracted with different polarities organic solvents with increasing polarities.The prepare hexane,chloroform,ethyl acetate,hutanol and methanol crude extracts were used for their evaluation of total phenol,flavonoids contents and phytochemical screening study.The established conventional metliods were used for quantitative determination of total phenol,flavonoids contents and phytochemical screening.Results:Phytochemical screening for various crude extracts were tested and shown positive result for flavonoids,saponins and steroids compounds.The result for total phenol content was the highest in hutanol and the lowest in methanol crude extract whereas the total flavonoids contents was the highest in methanol and the lowest hexane crude extract.Conclusions:The crude extracts from locally grown Thymus vulgaris showed high concentration of flavonoids and it could be used as antibiotics for different curable and uncurable diseases.

Extraction Process and Content Determination of Total Flavonoids in Ginkgo( Ginkgo biloba L. ) Leaves

[Objectives]This study was conducted to study the extraction process and the content determination of flavonoids in ginkgo( Ginkgo biloba L.) leaves.[Methods]Ethanol extraction and methanol extraction of total flavonoids in ginkgo leaves were studied,and the optimal extraction conditions for flavonoids were determined by orthogonal test; and with quercetin as reference substance,total flavonoid content in ginkgo leaves was determined by UV spectrophotometry.[Results]The optimal extraction process was 4 h of Soxhlet extraction with methanol; and the total flavonoid contents had a good linear relation in the range of 0. 006 5-0. 039 mg/ml( R^2= 0. 999 9),the average content was stabilized at 1. 135%,and the average recovery of the method was 102. 0%. [Conclusions]This study selected the optimal extraction process for total flavonoids in ginkgo leaves. The test method is simple with high accuracy and precision,and is suitable for the extraction and determination of total flavonoids in ginkgo leaves.

A new flavonoid from the bract of Zea mays L.

A new flavonoid was isolated from the bract of Zea mays L.The structure of the compound was identified as 4＇,5,7-trihydroxy-3＇,5＇-dimethoxyflavone 7-O-[β-D-apiofuranosyl（1→2）]-β-D-glucopyranoside on the ground of chemical and spectroscopic methods.

Extraction and Purification Process Optimization and Antioxidant Activity in vitro of Flavonoids in Amaranthus caudatus L.

[Objectives]To explore the optimal extraction and purification process of the flavonoids in Amaranthus caudatus L.and to study the antioxidant activity in vitro of the flavonoids in A.caudatus.[Methods]Taking A.caudatus as the raw material,flavonoids were extracted by alcohol extraction method,and AB-8 macroporous adsorption resin was selected for purification.The hydroxyl radical scavenging ability,DPPH radical scavenging ability,and O^2-radical scavenging ability were used as evaluation indicators,to explore the antioxidant activity in vitro of the flavonoids in A.caudatus.[Results]The optimal extraction process conditions of flavonoids in A.caudatus are:liquid-to-material ratio 40:1,extraction temperature 60℃,ethanol concentration 60%,ultrasonic power 320 W,extraction time 50 min.Under these conditions,the extraction yield of flavonoids in A.caudatus is(1.35±0.01)%.The optimal purification process conditions of flavonoids in A.caudatus are 2.5 g AB-8 macroporous adsorption resin,sample volume 5 mL,mass concentration of adsorption solution 1.60 mg/mL,pH value of adsorption solution 3.0,sample flow rate 3 BV/h,ethanol concentration in desorption process is 70%and the desorption flow rate is 3 BV/h.Under these conditions,the recovery rate reaches 88.35%±0.68%.[Conclusions]A.caudatus has a high content of flavonoids and has excellent free radical scavenging ability in vitro.This study is intended to provide important technical support for the research of flavonoid activity of A.caudatus and the development of functional products.

Effect of Total Flavonoids from Phyllanthus emblica L. on Tumor Proliferation and Immune Function

[Objectives]To explore the effect of total flavonoids from Phyllanthus emblica L.on tumor proliferation and immune function.[Methods]The effects of total flavonoids of P.emblica L.on the proliferation of 6 different tumor cell lines(human hepatoma cell line HepG-2,human cervical cancer cell line Hela,human gastric cancer cell line SGC7901,human nasopharyngeal carcinoma cell line CNE-2,human lung cancer cell line H460 and human ovarian cancer cell line A2780)were compared by SRB method.The effect of total flavonoids from P.emblica L.on the proliferation of mouse lymphocytes induced by concanavalin(ConA)or lipopolysaccharide(LPS)in vitro was detected by CCK-8 method.[Results]The results of SRB assay showed that compared with the normal group,the total flavonoids of P.emblica L.had obvious inhibitory effect on 6 kinds of tumor cells.Among them,the inhibitory effect on H460 cells and CNE-2 cells was the most significant,and the IC50 was(471.36±50.66),(463.26±40.75)μg/mL,respectively.The high dose group of total flavonoids from P.emblica L.had the same inhibitory effect as the positive drug 5-Fu.The results of CCK-8 assay showed that compared with the blank group,the total flavonoids of P.emblica L.significantly inhibited the proliferation of mouse lymphocytes induced by ConA or LPS(*P<0.05,**P<0.01).[Conclusions]The total flavonoids of P.emblica L.had significant anti-tumor activity.

Optimization on Ultrasonic Extraction Method of Total Flavonoids for Liriodendron chinense Sarg.×L.tulipifera L. by Response Surface Methodology (RSM)

With the leaves,bark and roots of Liriodendron chinense Sarg.× L.tulipifera L.as experiment materials,the ultrasonic-assisted extraction conditions of total flavonoids were optimized by response surface Box-Behnken test design.Ultrasonic time and liquid-to-solid ratio had significant effects on the extraction amount of flavonoids from the leaves of L.chinense Sarg.× L.tulipifera L.;ultrasonic temperature and liquid-to-solid ratio had an interaction on the extraction amount of flavonoids;and the optimum ultrasonic time,temperature and liquid-to-solid ratio for the extraction of flavonoids from leaves of L.chinense Sarg.× L.tulipifera L.were 19.82 min,28.60 ℃ and 9.48 ml/g,respectively.Ultrasonic time,temperature and liquid-to-solid ratio had significant effects on the extraction amount of flavonoids from L.chinense Sarg.× L.tulipifera L.;there was a significant interaction between any two of the factors,which had a significant effect on the extraction of flavonoids;and the optimum extraction parameters for flavonoids in the bark were the ultrasonic time of 33.66 min,the ultrasonic temperature at 32.85 ℃ and the liquid-to-solid ratio of 11.39 ml/g.Ultrasonic time and liquid-to-solid ratio had significant effects on the extraction of flavonoids from roots;there was a significant interaction between ultrasonic time and liquid-to-solid ratio and between ultrasonic temperature and liquid-to-solid ratio;and the optimum extraction parameters for flavonoids from the roots of L.chinense Sarg.× L.tulipifera L.were the ultrasonic time of 32.38 min,the ultrasonic temperature at 25.0 ℃ and the liquid-to-solid ratio of 8.00 ml/g.The results of the three models were stable.The models are feasible and have good application value.

Optimization of conditions for supercritical fluid extraction of flavonoids from hops (Humulus lupulus L.)

Waste hops are good sources of flavonoids. Extraction of flavonoids from waste hops (SC-CO2 extracted hops) using supercritical fluids technology was investigated. Various temperatures, pressures and concentrations of ethanol (modifier) and the ratio (w/w) of solvent to material were tested in this study. The results of single factor and orthogonal experiments showed that at 50 °C, 25 MPa, the ratio of solvent to material (50%), ethanol concentration (80%) resulted in maximum extraction yield fla- vonoids (7.8 mg/g). HPLC-MS analysis of the extracts indicated that flavonoids obtained were xanthohumol, the principal prenylflavonoid in hops.

UPLC-Q-Exactive Orbitrap-MS法分析光果甘草地上部分黄酮类成分

[目的]对光果甘草地上部分(茎、叶)的黄酮类化学成分进行分析,为进一步开发利用光果甘草地上部分及阐明其药效物质基础提供科学依据。[方法]将光果甘草地上部分醇提样品水沉,分为水溶性、脂溶性部位;采用超高效液相色谱-四级杆-静电场轨道阱联用(UPLC-Q-Exactive Orbitrap-MS)法检测样品,使用Xcalibur软件处理分析数据,将得到的成分分子质量、多级图谱碎片信息结合文献、数据库及标准品进行指认。[结果]从光果甘草地上部分共检测出100个黄酮类成分,有97种在光果甘草地上部分被首次发现;其中水溶性样品检测出61种,脂溶性样品检测出69种。[结论]通过UPLC-Q-Exactive Orbitrap-MS法,对光果甘草地上部分建立了快速、准确的定性方法,同时证实黄酮类成分含量丰富,为进一步开发利用提供依据。